Summary of Proposed Work - A DNA-directed system for RNA and protein synthesis is being used to quantitatively analyze the factors necessary for regulation of genes associated with the lac and ara operons, the su3 ion tRNA, Co1E1 and related hybrid plasmids. The factors which are being studied in this synthetic system or in more purified systems include the catabolite gene activator protein, CAP, the ara C protein and magic spot, ppGpp. Radioactively labeled RNAs and proteins synthesized in the cell-free system are being characterized primarily by electrophoresis on gels and fluorography.